Method of enhancing the host defense

ABSTRACT

The invention relates to a method of enhancing or regulating the host defense of a mammal, said method comprising administering to a mammal a therapeutically effective amount of hyaluronic acid.

This is a continuation of U.S. application Ser. No. 726,956, filed Apr.26, 1985, which is a continuation of application Ser. No. 455,845, filedJan. 5, 1983, which is a continuation of application Ser. No. 261,800,filed May 8, 1981, which is a continuation of application Ser. No.045,599, filed June 5, 1979, now all abandoned.

The present invention relates to a method of stimulating or regulatingthe host defence in mammals, i.e. to fortify a mammal's resistance toinfections.

In many instances it is desirable to stimulate and enhance theimmunological and related systems (herein referred to as the "hostdefence") of mammals, especially humans, to increase the resistance toinfections diseases. This would not only be desirable for patients witha reduced host defence, but also, in many instances, for patients havinga normal host defence. For example, reduced resistance to infectiousdiseases is a common clinical problem (involving both primary andsecondary defects) appearing e.g. with severly burnt patients, patientswith malignancies receiving cytotoxic drugs, uremic patients, diabetics,patients suffering from chronic staphylococcus disease, chronicbronchitis, and so on. Examples of conditions where regulation of anormal host defence is desirable are e.g. immunecomplex diseases such asSLE (systemic lupus erthematosus, etc., granulomatous diseases such aslepra, sarcoidosis, Crohn's disease, etc., malignancies, and the like.

Thus there is an urgent need for means and methods to make it possibleto enhance or stimulate the host defence in mammals. However, no suchmeans or methods are available at present.

It is an object of the present invention to solve this problem and toprovide a method of enhancing or regulating the host defence in mammals.

It is another object of the invention to provide a method of preventing,inhibiting or curing diseases involving an insufficient host defence,especially the above mentioned and similar diseases.

The present invention is based on the unexpected discovery thatadministration of hyaluronic acid (which is a known substance) tomammals results in a considerable increase of the host defence. Thiseffect has been confirmed by in vivo tests, as will be explained more indetail below.

The hyaluronic acid is preferably administered parenterally especiallysubcutaneously and also intra-muscularly or intra-venously. Forparenteral administration the hyaluronic acid is preferably used in theform of a solution of a physiologically acceptable salt thereof in aphysiological buffer..

The hyaluronic acid is preferably administered in doses from 1 to 40 mg,especially from 5 to 20 mg. The effect of the hyaluronic acid on thehost defence usually has a rather extended duration (e.g. about one weekor more), so in some cases one single administration is sufficient. Inother cases repeated administrations are required, e.g. oneadministration each week. The suitable dose and frequency ofadministration can conveniently be determined in each individual case bytaking blood samples from the patient for monitoring the level of hostdefense stimulation, e.g. by means of the procedures described below,and adjusting the dose level and administration frequency in responsethereto. The treatment with hyaluronic acid in accordance with theinvention has proved to be virtually without any undesired side effects.

One type of hyaluronic acid, which is suitable for the purposes of thepresent invention, is the essentially pur hyaluronic acid disclosed inU.S. Pat. No. 4,141,973, which is incorporated by reference herein. Thishyaluronic acid has the following characteristics: an average molecularweight of at least about 750,000, a protein content of less than 0.5% byweight, ultraviolet light absorbance of a 1% solution of the sodium saltthereof of less than 3.0 at 257 nanometers wavelength and less than 2.0at 280 nanometers wavelength, a kinematic viscosity of a 1% solution ofthe sodium salt thereof in physiological buffer of greater than about1000 centistokes and a molar optical rotation of a 0.1-0.2% solution ofthe sodium salt thereof in physiological buffer of less than -11×10³degree-cm² /mole (of disaccharide) measured at 220 nanometers. Furthercharacteristics of said hyaluronic acid fraction, which is sterile,pyrogen-free, protein-free and non-antigenic, are the absence ofsignificant cellular infiltration of the vitreous and anterior chamber,absence of significant flare in the aqueous humor, absence ofsignificant haze or flare in the vitreous and absence of pathologicalchanges to the cornea, lens, iris, retina, and choroid of the owl monkeyeye when on millileter of a 1% solution of the sodium salt thereofdissolved in physiological buffer is implanted in the vitreous replacingabout one-half of the existing liquid vitreous. However, the inventionis not intended to be restricted to the use of this specific hyaluronicacid, but any type of hyaluronic acid can be used, which is capable ofcausing an enhanced host response when administered to mammals inaccordance with this invention.

The invention will be described more in detail in the followingnonlimiting examples relating to the treatment of a plurality ofpatients having increased infection propensity with hyaluronic acid inaccordance with the present invention. The preparation used in theseExamples was Healon®, provided by Pharmacia AB, Uppsala, Sweden. Thispreparation was an aqueous sterile solution for injection, 1 ml of whichcontaining:

Sodium hyaluronate: 10 mg

Sodium chloride: 8.5 mg

Sodium dihydrogen phosphate hydrate: 0.040 mg

Disodium hydrogen phosphate dihydrate: 0.28 mg

Water: q.s.

This preparation is hereinafter referred to as "HA". The sodiumhyaluronate had an average molecular weight of 1.5×10⁶, a limitingviscosity of 4000 ml/g and a protein content of 0.15%. The stimulationof the host defence (neutrophil function) was monitored by kineticstudies of the phagocytic uptake of IgG or complement-coated latexparticles according to the principles outlined in Hallgren et al,"Kinetic Studies of Phagocytosis of IgG-coated Latex Particles withThrombocyte Counter", Journal of Laboratory and Clinical Medicine, Vol.90, No. 5, pp 786-795, which is incorporated by reference herein.

EXAMPLE 1 Woman, Age 57 Years

Clinical diagnosis and symptoms before the treatment. The patient has achronic leg wound and a strongly increased propensity to infections. Shehas, during the last 5 to 6 years, repeatedly suffered from bacterialinfections of the meningitis, sepsis type. Since autumn 1977 she hasbeen confined to bed with a febris continua, which has now and then beeninterrupted by septical fever peeks.

Phagocytosis function before the treatment. The patient had a normalphagocytosis of IgG opsonized particles, but a strongly defectiveability to phagocytize serum opsonized particles. This points to adisturbance of the membrane receptor function of the neutrophil.

Treatment. The patient was given 1 ml of HA subcutaneously on thefollowing occasions: 1 dose each week from June 13, 1978 to Oct. 1,1979. The treatment was continued on Nov. 23, 1978 with 1 ml of HAsubcutaneously ever week and was still going on (May 15, 1979).

Development after the treatment. The patient was treated and observedfor 3 weeks as a patient at the University Hospital, Uppsala, Sweden.During this period the patient was still confined to bed, but the feverperiods declined somewhat. The treatment was continued during the summerand she became substantiallyfree from infections, could be mobilized andhad been at home. The treatment was continued until Oct. 1, 1978, whenthe patient herself requested the same to be discontinued. The effect onthe leg wound had been somewhat uncertain, but the patient had no feverand was no longer confined to bed. About 3 weeks after the lastinjection the fever peeks of the patient became more and more frequent,and the patient requested continued treatment as she had felt betterduring the first period of treatment. The patient was once againhospitalized at the University Hospital, the phagocytosis function wasevaluated and the treatment with hyaluronic acid was started again.About 10 days after the first injection the fever periods of the patientwere once again reduced. The patient has returned to her home town andher general condition has been considerably improved. The leg wound,which has been chronic for about 6 to 7 years, is now healing.

Phagocytosis function after treatment. The granulocyte function has beennormalized during the treatment, meaning that the IgG uptake is stillgood and that the serum dependent particle uptake has been completelynormalized.

Side effects. No side effects have been discovered, except for minorskin bleedings around the points of injection.

Conclusion. This is a very ill patient with a long and serious infectionanamnesis and confinement to bed for almost 1 year due to seriousinfections. In connection with the treatment with hyaluronic acid thepatient could be mobilized and became free of fever. When the treatmentwas interrupted the fever periods returned. Resumption of treatment withhyaluronic acid resulted in an improvement in the patient's functionsubjectively and objectively.

EXAMPLE 2 Man, Age 53 Years

Clinical diagnosis and symptoms before treatment. The patient is anextremely infection sensitive individual, who has been examined andtreated at various departments of the University Hospital for severalyears. Among other things, he has a strange lung infiltration, whichsometimes has been interpreted as sarcoidosis and sometimes as a chronicfungal infection. The dominating symptoms of the patient have been veryfrequent bacterial infections with fever periods. Furthermore, he hasdeveloped a heart insufficiency due to cardial valve defects, andaccording to thorax surgical experts and cardiological experts thepatient required an operation in order not to die of this disease.However, this has not been done because of the abnormal infectionpropensity of the patient.

Phagocytosis function before treatment. The patient has a reduced uptakeboth of IgG and serum opsonized particles.

Treatment. The patient has been given 1 ml of HA subcutaneously on thefollowing occasions: Dec. 19 and 31, 1978 and Feb. 6 and 13, 1979.

Development after treatment. The patient was operated on Dec. 18, 1978for the valve defect, and the post-operative development was completelyfree from infections. The patient returned to the hospital Mar. 5, 1979since he had again had a bacterial infection, this time a sinusitis. Byexperience the patient's infections have been hard to treat, and thepatient was therefore once again given hyaluronic acid in combinationwith an antibiotic. The condition of the patient was rapidly improved.

Phagocytosis function after treatment. Completely normalized.

Side effects. No side effects were observed.

Conclusion. This is an abnormally infectionprone individual, who hasbeen tested by us several times during the last years. Because of thesevere heart valve defect it was considered vital to operate on thepatient. Hyaluronic acid was given in order to prevent the possibledevelopment of infections in the post-operative stage. The patient hadno post-operative complications.

EXAMPLE 3 Woman, Age 37 Years

Clinical diagnosis and symptoms before treatment. The patient has achronic bronchitis with emphysema. According to lung specialists theprognosis was ver pessimistic. The patient has had repeated pneumonicinfections with fever, about 3 to 4 per month.

Phagocytosis function before treatment. Reduced phagocytosis of both IgGparticles and serum opsonized particles.

Treatment. 1 ml of HA subcutaneously once every week. The treatmentstarted Jan. 26, 1979 and was still continued on May 15, 1979.

Development after treatment. After one month of treatment the patientand her husband have experienced a subjective improvement. Compared to 3to 4 infection periods per month the patient has during the last threemonths only had 1 infection episode per month.

Phagocytosis function after treatment. A considerably improvedgranulogyte function both with regard to the uptake of IgG and serumopsonized particles.

Side effects. No side effects observed except for a slight tenderness atthe points of injection.

Conclusion. A patient with a severe bronchitis, emphysema developmentand a pessimistic prognosis quo ad vitam has been treated for a littlemore than one month with hyaluronic acid, and she has during this periodexperienced a clear improvement with regard to the number of infectionepisodes.

EXAMPLE 4 Man, Age 36 Years

Clinical diagnosis and symptoms before treatment. the patient had skinburns on both arms, caused by electricity. The total area of the burntskin was 9%. Due to their total distruction the patient's arms wereamputated. The patient had severe infections in the amputation areas.

Phagocytosis function before treatment. The phagocytosis function of thepatient was monitored for two weeks. It was progressively worsened andat the end of the two week period, the phagocytosis function was verypoor.

Treatment. 1 ml of HA subcutaneously on Oct. 10, 1978 and 2 ml of HAsubcutaneously on Oct. 18, 1978.

Development after treatment. The general condition of the patient wasvery poor in relation to the size of the skin burns. The patient was ina respirator before the treatment and was dialyzed peritoneally. Inconnection with the treatment the patient's infected areas dried up andhe could be taken out from the respirator after only a few days. Thegeneral condition was remarkably improved during the following 2 weeksand the patient was sent home 3 weeks later.

Phagocytosis function after treatment. The phagocytosis function rosesomewhat after the first 1 ml injection, but then returned to the samelow level. A strong increase of the uptake of both IgG and serumopsonized particles was observed after the 2 ml injection.

Side effects. None observed.

EXAMPLE 5 Man, Age 11 Years

Clinical diagnosis and symptoms before treatment. A patient with a skinburn covering 37% of the skin area. Amputation of one arm and both legswas necessary due to extended necrosis. The patient was treated in arespirator and was unconscious from the very beginning and showedsymptoms of a slight cerebral atrophy.

Phagocytosis function before treatment. the phagocytosis function wasfollowed for one week. An increasing reduction to very low levels of theuptake of both serum opsonized and IgG particles was observed.

Treatment. 1 ml of HA subcutaneously Dec. 15, 1978, 0.5 ml of HAsubcutaneously Jan. 4, 1979, 1 ml of HA subcutaneously Jan. 18, 1979.

Development after treatment. The large wound areas of the patient driedup somewhat. After a while the patient woke up to consciousness andcould be taken out from the respirator. He was sent to his home townhospital on Feb. 2, 1979.

Phagocytosis function after treatment. Phagocytosis of both the IgG andserum opsonized particles increased in connection with the treatmentsand the improvement remained for about one week after the injection.

Side effects. None observed.

Conclusion. A severely burned boy had been subjected to high tensionelectricity. The general condition was poor, as was the phagocytosisfunction before the treatment with the hyaluronic acid. In connectionwith this treatment the phagocytosis function was improved and the woundareas of the patient dried up. He could be sent home after about 2months at the University Hospital.

EXAMPLE 6 Man, Age 48 Years

Clinical diagnosis and symptoms before treatment. A patient having a 25%skin burn with an initially very poor general condition and extendedskin infections was studied.

Phagocytosis function before treatment. The patient had at the beginninga strong reduction of the uptake of IgG opsonized particles. The serumopsonized particle uptake diminishes progressively and after tow weekswas strongly reduced.

Treatment. 1 ml of HA subcutaneously on Feb. 9 and 16, 1979.

Development after treatment. The general condition of the patientimproved very rapidly during and after the treatment. He was sent homeon Mar. 7, 1979.

Phagocytosis function after treatment. A considerable improvement of thephagocytosis function after each treatment, especially with regard tothe uptake of serum opsonized particles was observed.

Side effects. None observed.

Conclusion. The patient had an average size skin burn and infectioncomplication in the wound areas. He initially had a very poor generalcondition and a poor phagocytosis function. Both parts were improved inconnection with the treatment with hyaluronic acid.

What we claim is:
 1. A method for normalizing a reduced phagocyticactivity exerted by the granulocytes of a mammal, comprisingadministering subcutaneously or intramuscularly to said mammal anon-antigenic hyaluronic acid preparation containing hyaluronic acid ora physiologically acceptable salt of said acid in a therapeuticallyeffective amount.
 2. The method according to claim 1, wherein saideffective amount comprises 1-40 mg of hyaluronic acid or aphysiologically acceptable salt thereof.
 3. A method for treating orpreventing infections associated with reduced phagocytic activity, inmammals, comprising administering subcutaneously or intramuscularly to amammal a therapeutically effective amount of a non-antigenic hyaluronicacid preparation containing hyaluronic acid or a physiologicallyacceptable salt thereof.
 4. The method according to claim 3, whereinsaid effective amount comprises 1-40 mg of hyaluonic acid or aphysiologically acceptable salt thereof.
 5. The method according toclaim 3, wherein the mammal to be treated exhibits reduced granulocytephagocytic activity.
 6. The method according to claim 4, wherein themammal to be treated exhibits reduced granulocyte phagocytic activity.7. The method claim 1, wherein the mammal is a human.
 8. The method ofclaim 3, wherein said mammal is a human.
 9. A method for treatinginfections in humans associated with reduced phagocytic activity,comprising administering subcutaneously or intramuscularly to a human atherapeutically effective amount of a non-antigenic hyaluronic acid or aphysiologically acceptable salt thereof.
 10. The method according toclaim 9, wherein the human to be treated exhibits reduced granulocyticactivity.
 11. The method according to claim 9, wherein said effectiveamount comprises 1-40 mg of hyaluronic acid or a physiologicallyacceptable salt thereof.
 12. The method according to claim 11, whereinthe human to be treated exhibits reduced granulocytic activity.